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Immunohistological expression of cytochrome P450 1A2 (CYP1A2) in the ovarian follicles of prepubertal and pubertal rat

  • Hwang, Jong-Chan;Park, Byung-Joon;Kim, Hwan-Deuk;Baek, Su-Min;Lee, Seoung-Woo;Jeon, Ryoung-Hoon;Jang, Min;Bae, Seul-Gi;Yun, Sung-Ho;Park, Jin-Kyu;Kwon, Young-Sam;Kim, Seung-Joon;Lee, Won-Jae
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.4
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    • pp.329-337
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    • 2020
  • Cytochrome P450 1A2 (CYP1A2) is a member of the cytochrome P450 superfamily enzymes in mammals and plays a major role in metabolizing endogenous hormones in the liver. In recent days, CYP1A2 expression has been found in not only the liver but also other tissues including the pancreas and lung. However, little information is available regarding the expression of CYP1A2 in the ovary, in spite of the facts that the ovarian follicle growth and atresia are tightly associated with controls of endocrine hormonal networks. Therefore, the expression of CYP1A2 in the ovaries of prepubertal and pubertal rats was investigated to assess its expression pattern and puberty-related alteration. It was demonstrated that the expression level of CYP1A2 was significantly (p < 0.01) higher in the pubertal ovaries than prepubertal counterparts. At the ovarian follicle level in both groups, whereas CYP1A2 expression was less detectable in the primordial, primary and secondary follicles, the strongly positive expression of CYP1A2 was localized in the granulosa cell layers in the antral and pre-ovulatory follicles. However, the ratio of CYP1A2-positive ovarian follicle was significantly (p < 0.01) higher in the ovary of pubertal group (73.1 ± 3.1%) than prepubertal one (41.0 ± 10.5%). During the Immunofluorescence, expression of CYP1A2 was mainly localized in Fas-positive follicles, indicating the atretic follicles. In conclusion, these results suggested that CYP1A2 expression was mainly localized at the atretic follicular cells and affected by the onset of puberty. Further study is still necessary but we hypothesize that CYP1A2 expresses in the atretic follicles to metabolize residue of the reproductive hormones. These findings may have important implications for the fields of reproductive biology of animals.

Molecular Analysis of the Interaction between Human PTPN21 and the Oncoprotein E7 from Human Papillomavirus Genotype 18

  • Lee, Hye Seon;Kim, Min Wook;Jin, Kyeong Sik;Shin, Ho-Chul;Kim, Won Kon;Lee, Sang Chul;Kim, Seung Jun;Lee, Eun-Woo;Ku, Bonsu
    • Molecules and Cells
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    • v.44 no.1
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    • pp.26-37
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    • 2021
  • Human papillomaviruses (HPVs) cause cellular hyperproliferation-associated abnormalities including cervical cancer. The HPV genome encodes two major viral oncoproteins, E6 and E7, which recruit various host proteins by direct interaction for proteasomal degradation. Recently, we reported the structure of HPV18 E7 conserved region 3 (CR3) bound to the protein tyrosine phosphatase (PTP) domain of PTPN14, a well-defined tumor suppressor, and found that this intermolecular interaction plays a key role in E7-driven transformation and tumorigenesis. In this study, we carried out a molecular analysis of the interaction between CR3 of HPV18 E7 and the PTP domain of PTPN21, a PTP protein that shares high sequence homology with PTPN14 but is putatively oncogenic rather than tumor-suppressive. Through the combined use of biochemical tools, we verified that HPV18 E7 and PTPN21 form a 2:2 complex, with a dissociation constant of 5 nM and a nearly identical binding manner with the HPV18 E7 and PTPN14 complex. Nevertheless, despite the structural similarities, the biological consequences of the E7 interaction were found to differ between the two PTP proteins. Unlike PTPN14, PTPN21 did not appear to be subjected to proteasomal degradation in HPV18-positive HeLa cervical cancer cells. Moreover, knockdown of PTPN21 led to retardation of the migration/invasion of HeLa cells and HPV18 E7-expressing HaCaT keratinocytes, which reflects its protumor activity. In conclusion, the associations of the viral oncoprotein E7 with PTPN14 and PTPN21 are similar at the molecular level but play different physiological roles.

Rapid Cell Death Phenotype of Streptococcus mutans under Prolonged Growth Conditions (장시간 생장 조건에서 Streptococcus mutans의 급격한 세포사 표현형 분석)

  • Kim, Jeong Nam
    • Journal of Life Science
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    • v.31 no.12
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    • pp.1072-1078
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    • 2021
  • The oral pathogen Streptococcus mutans is considered a major causative agent of dental caries in humans. The use of dental hygiene products, including toothpaste and mouthwash, is used for caries control. However, food intake can lead to the recurrence of oral microorganisms. This study aimed to explore why this bacterium dies so quickly during prolonged incubation and to assess whether this growth characteristic is closely associated with the secretion of metabolic products. Notably, the number of live S. mutans cells rapidly declined after 24 hr during the entire period tested, whereas the number of Escherichia coli cells, an indicator strain, remained steady over the same period. To test whether the S. mutans supernatants contained possible signals that accelerated the death of neighbor cells, we obtained the individual supernatants at the above time points. Following pH neutralization, the cells in which the supernatant was supplemented with glucose grew well. However, pH adjustment alone could not fully recover cell growth in conditions in which the supernatant was supplemented, with or without glucose. These phenotypes of S. mutans may be associated with signaling, not only resulting from nutrient depletion. The findings on the survival phenotype of S. mutans provide new insights into cell-cell communication in the biology of this bacterium.

Biochemical Adaptation of the Oriental Tobacco Budworm, Helicoverpa assulta, to Host-plant Defensive Compounds (기주식물 방어물질에 대한 담배나방의 생화학적 적응)

  • Ahn, Seung-Joon
    • Korean journal of applied entomology
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    • v.61 no.1
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    • pp.143-154
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    • 2022
  • Plant secondary metabolites play an important role in insect-plant interactions. Herbivorous insects have various strategies to cope with the plant defensive compounds. Polyphagous insects feed on a wide variety of plant species, and their detoxification mechanisms are more complex since they tend to respond to a large array of different plant-derived chemicals. Alternatively, oligophagous insects specialize on only a few related plant species and may be expected to have a more efficient form of adaptation. This adaptation could involve either the production of large quantities of enzymes to detoxify their defensive compounds or the sequestration of the compounds or their metabolites. The oriental tobacco budworm, Helicoverpa assulta, is a specialist herbivore, feeding on a few plants of Solanaceae, such as tobacco and hot pepper. Understanding its host-plant adaptation not provides an important insight on physiology, ecology and evolution of specialist herbivores, but also gives a clue to develop management strategies of the pest species such as H. assulta. This paper briefly reviews the specialist, H. assulta, focusing on its host range, larval associations with the host plants, and detoxification mechanisms to nicotine and capsaicin, two characteristic defensive compounds derived from its two major host plants, tobacco and hot pepper, respectively. It summarizes the relevant research over the last half century and provides a future perspective on this subject.

Changes in the Chlorophyll of Garlic Chives (Allium tuberosum) Resulting fromFertilizer and Drought Stress (비료와 가뭄 스트레스에 의한 부추의 엽록소 변화)

  • Huh, Man Kyu;Lee, Byeongryong
    • Journal of Life Science
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    • v.32 no.10
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    • pp.743-748
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    • 2022
  • The garlic chive (Allium tuberosum Rotter) is a prominent herb species in Asia and other nations of the world. Garlic chives is a favorite vegetable and used to garnish noodles in Korea. The effects of various doses of N, P, and K fertilizers and drought stress on the chlorophyll content in the leaves of garlic chives were investigated. The evaluations showed that chlorophyll a content was 0.386 at 10 mg/l N fertilizer and 0.584 at 50 mg/l N fertilizer. The treatment group showed a significant difference with regard to the contents of chlorophylls a and b and total chlorophyll at the 5% level (p<0.05). Pearson's correlation coefficient (Pearson's r) for chlorophylls a and b and total chlorophyll were 0.940, 0.966, and 0.971, respectively. The highest content of chlorophylls a and b and total chlorophyll in the leaves was recorded at 40 mg/l P fertilizer, while the values corresponding to 50 mg/l P fertilizer were lower than those for 40 mg/l P fertilizer. The content of total chlorophyll evaluated at 10 mg/l K fertilizer was 0.312 and that at 50 mg/l was 0.589. The simple linear regression showed the relationship between chlorophyll efficiency aand moisture. The slope factors of the dark-level fluorescence yield (Fo), the maximum fluorescence yield (Fm), the quenched state (Fv), and the maximal PSII quantum yield (Fv/Fm) for chlorophyll-efficient indicators were -0.931, 0.972, 972, and 0.950, respectively. NPK fertilizers and drought stress affected the chlorophyll content and efficiency of A. tuberosum.

Regulation of tumor-associated macrophage (TAM) differentiation by NDRG2 expression in breast cancer cells

  • Lee, Soyeon;Lee, Aram;Lim, Jihyun;Lim, Jong-Seok
    • BMB Reports
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    • v.55 no.2
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    • pp.81-86
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    • 2022
  • Macrophages are a major cellular component of innate immunity and are mainly known to have phagocytic activity. In the tumor microenvironment (TME), they can be differentiated into tumor-associated macrophages (TAMs). As the most abundant immune cells in the TME, TAMs promote tumor progression by enhancing angiogenesis, suppressing T cells and increasing immunosuppressive cytokine production. N-myc downstream-regulated gene 2 (NDRG2) is a tumor suppressor gene, whose expression is down-regulated in various cancers. However, the effect of NDRG2 on the differentiation of macrophages into TAMs in breast cancer remains elusive. In this study, we investigated the effect of NDRG2 expression in breast cancer cells on the differentiation of macrophages into TAMs. Compared to tumor cell-conditioned medium (TCCM) from 4T1-mock cells, TCCM from NDRG2-over-expressing 4T1 mouse breast cancer cells did not significantly change the morphology of RAW 264.7 cells. However, TCCM from 4T1-NDRG2 cells reduced the mRNA levels of TAM-related genes, including MR1, IL-10, ARG1 and iNOS, in RAW 264.7 cells. In addition, TCCM from 4T1-NDRG2 cells reduced the expression of TAM-related surface markers, such as CD206, in peritoneal macrophages (PEM). The mRNA expression of TAM-related genes, including IL-10, YM1, FIZZ1, MR1, ARG1 and iNOS, was also downregulated by TCCM from 4T1-NDRG2 cells. Remarkably, TCCM from 4T1-NDRG2 cells reduced the expression of PD-L1 and Fra-1 as well as the production of GM-CSF, IL-10 and ROS, leading to the attenuation of T cell-inhibitory activity of PEM. These data showed that compared with TCCM from 4T1-mock cells, TCCM from 4T1-NDRG2 cells suppressed the TAM differentiation and activation. Collectively, these results suggest that NDRG2 expression in breast cancer may reduce the differentiation of macrophages into TAMs in the TME.

Pathophysiological Functions of Deubiquitinating Enzymes in Obesity and Related Metabolic Diseases (탈유비퀴틴화 효소 DUBs의 비만 및 대사 관련 질환에서 병태생리학적 기능)

  • Lee, Seul Gi;Kwon, Taeg Kyu
    • Journal of Life Science
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    • v.32 no.6
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    • pp.476-481
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    • 2022
  • Ubiquitin signaling regulates virtually all aspects of eukaryotic biology and dynamic processes in which protein substrates are modified by ubiquitin. To regulate these processes, deubiquitinating enzymes (DUBs) cleave ubiquitin or ubiquitin-like proteins from these substrates. DUBs have been implicated in the pathogenesis of cancer, leading to the development of increasing numbers of small-molecule DUB inhibitors. On the other hand, recent studies have focused on the function of DUBs in metabolic diseases such as obesity, diabetes, and fatty liver diseases. DUBs play a positive or negative role in the progression and development of metabolic diseases. Their involvement in cell pathology and regulation of major transcription factors in metabolic syndrome has been examined in vitro and in animal and human biopsies. UCH, USP7, and USP19 were linked to adipocyte differentiation, body weight gain, and insulin resistance in genetic or diet-induced obesity. CYLD, USP4, and USP18 were found to be closely associated with fatty liver diseases. In addition, these liver diseases were accompanied by body weight change in certain cases. Collectively, in this review, we discuss the current understanding of DUBs in metabolic diseases with a particular focus on obesity. We also provide basic knowledge and regulatory mechanisms of DUBs and suggest these enzymes as therapeutic targets for metabolic diseases.

Test of Superoxide Dismutase Characteristics and Antioxidant Activity in Perilla Leaves (들깨잎에 함유된 Superoxide Dismutase의 특성 및 항산화 활성 검정)

  • Chung, Ill-Min;Yun, Song-Joong;Kim, Jung-Tae;Gwag, Jae-Gyun;Sung, Jae-Duck;Suh, Hyung-Soo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.4
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    • pp.504-511
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    • 1995
  • This study was conducted to investigate the superoxide dismutase (SOD) characteristics and antioxidant activity by nonenzymatic(Fe$^{2+}$/Ascorbate) and Fe$^{3+}$-ADP/NADPH method in perilla(Perilla frutescens var. japonica Hara.) and jaso(Perilla frutescens Briton var. acuta Kudo.) leaves. The characteristics were evaluated by the nitro blue tetrazolium reduction method. Perilla leaves contained three or four major SODs depending on the varieties. The inhibitor test indicated that the Perilla leaves contained two Cu /ZnSODs and one or two FeSODs, but Jaso leaves have only Cu/ZnSOD. However, no varietal differences were detected in the Cu /ZnSOD isozyme patterns. FeSODs, however, showed different varietal isozyme patterns through the different combinations of the two FeSOD isozymes. Among MeOH extractes, "mil yang 2" showed very strong antioxidant activity. Relatively large differences in the levels of SOD and antioxidant activity detected in the Perilla varietites. There was significantly different in the comparison between perilla leaves and red jaso leaves.s.etween perilla leaves and red jaso leaves.

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Mountain-cultivated ginseng protects against cognitive impairments in aged GPx-1 knockout mice via activation of Nrf2/ChAT/ERK signaling pathway

  • Bao Trong Nguyen;Eun-Joo Shin;Ji Hoon Jeong;Naveen Sharma;Ngoc Kim Cuong Tran;Yen Nhi Doan Nguyen;Dae-Joong Kim;Myung Bok Wie;Yi Lee;Jae Kyung Byun;Sung Kwon Ko;Seung-Yeol Nah;Hyoung-Chun Kim
    • Journal of Ginseng Research
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    • v.47 no.4
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    • pp.561-571
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    • 2023
  • Background: Escalating evidence shows that ginseng possesses an antiaging potential with cognitive enhancing activity. As mountain cultivated ginseng (MCG) is cultivated without agricultural chemicals, MCG has emerged as a popular herb medicine. However, little is known about the MCG-mediated pharmacological mechanism on brain aging. Methods: As we demonstrated that glutathione peroxidase (GPx) is important for enhancing memory function in the animal model of aging, we investigated the role of MCG as a GPx inducer using GPx-1 (a major type of GPx) knockout (KO) mice. We assessed whether MCG modulates redox and cholinergic parameters, and memory function in aged GPx-1 knockout KOmice. Results: Redox burden of aged GPx-1 KO mice was more evident than that of aged wild-type (WT) mice. Alteration of Nrf2 DNA binding activity appeared to be more evident than that of NFκB DNA binding activity in aged GPx-1 KO mice. Alteration in choline acetyltransferase (ChAT) activity was more evident than that in acetylcholine esterase activity. MCG significantly attenuated reductions in Nrf2 system and ChAT level. MCG significantly enhanced the co-localization of Nrf2-immunoreactivity and ChAT-immunoreactivity in the same cell population. Nrf2 inhibitor brusatol significantly counteracted MCG-mediated up-regulation in ChAT level and ChAT inhibition (by k252a) significantly reduced ERK phosphorylation by MCG, suggesting that MCG might require signal cascade of Nrf2/ChAT/ERK to enhance cognition. Conclusion: GPx-1 depletion might be a prerequisite for cognitive impairment in aged animals. MCG-mediated cognition enhancement might be associated with the activations of Nrf2, ChAT, and ERK signaling cascade.

Big Data Analytics in RNA-sequencing (RNA 시퀀싱 기법으로 생성된 빅데이터 분석)

  • Sung-Hun WOO;Byung Chul JUNG
    • Korean Journal of Clinical Laboratory Science
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    • v.55 no.4
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    • pp.235-243
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    • 2023
  • As next-generation sequencing has been developed and used widely, RNA-sequencing (RNA-seq) has rapidly emerged as the first choice of tools to validate global transcriptome profiling. With the significant advances in RNA-seq, various types of RNA-seq have evolved in conjunction with the progress in bioinformatic tools. On the other hand, it is difficult to interpret the complex data underlying the biological meaning without a general understanding of the types of RNA-seq and bioinformatic approaches. In this regard, this paper discusses the two main sections of RNA-seq. First, two major variants of RNA-seq are described and compared with the standard RNA-seq. This provides insights into which RNA-seq method is most appropriate for their research. Second, the most widely used RNA-seq data analyses are discussed: (1) exploratory data analysis and (2) pathway enrichment analysis. This paper introduces the most widely used exploratory data analysis for RNA-seq, such as principal component analysis, heatmap, and volcano plot, which can provide the overall trends in the dataset. The pathway enrichment analysis section introduces three generations of pathway enrichment analysis and how they generate enriched pathways with the RNA-seq dataset.